• CHANDANA CHOUDHURY BARUA Department of Pharmacology and Toxicology, College of Veterinary Science, Assam Agricultural University, Guwahati, Assam 781022, India
  • S. M. ABDUL AZIZ BARBHUIYA Department of Pharmacology and Toxicology, College of Veterinary Science, Assam Agricultural University, Guwahati, Assam 781022, India
  • LIPIKA BURAGOHAIN Department of Pharmacology and Toxicology, College of Veterinary Science, Assam Agricultural University, Guwahati, Assam 781022, India
  • AJAY KAKATI Department of Pharmacology and Toxicology, College of Veterinary Science, Assam Agricultural University, Guwahati, Assam 781022, India
  • ACHEENTA GOHAIN BARUA Department of Veterinary Public Health, College of Veterinary Science, Assam Agricultural University, Guwahati, Assam 781022, India


Objective: To investigate the analgesic activity of methanolic extract of Entada phaseoloides (MEEP) along with its molecular mechanistic pathway.

Methods: Swiss albino female mice and Wistar rats of either sex were administered orally with MEEP extracts (100, 200 and 400 mg/kg) and pentazocine, tramadol and diclofenac sodium, as standard drugs. Following administration, anti-nociceptive activity was evaluated using an acetic acid-induced writhing test, Eddy’s hot plate, and hot water immersion test. Serum was collected for molecular expression of various proteins and genes using Reverse Transcriptase PCR and Western Blotting.

Results: Acetic acid writhing test, a frequently used method to assess peripheral analgesic activity, revealed that MEEP reduced peripherally induced pain in a dose-dependent manner. Likewise, Eddy’s hot plate and hot water immersion methods, often implicated for testing central analgesic activities, showed that MEEP is bestowed with the capability to counteract analgesia in a dose and time-dependent manner. Pro-inflammatory cytokines and factors like COX-2, IL-6, TNF-α, and NF-κB that cause inflammatory responses and pain were significantly reduced, suggesting its analgesic and anti-inflammatory potential. This analgesic and the anti-inflammatory role played by MEEP is also supported by the up-regulation of anti-inflammatory cytokine IL-10 and COMT and COX-1 enzyme demonstrated no significant difference between the groups.

Conclusion: The study revealed the weak peripheral and potent central analgesic property MEEP by modulating pro-inflammatory and anti-inflammatory pathways.

Keywords: Analgesic, Cytokines, Diclofenac sodium, Entada phaseoloides, Pentazocine, Tramadol


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1. Kirtikar KR, Basu BD, Blatter E. 4th Vol. 2nd edition. Indian Medicinal Plants. Lalit Mohan Basu Publishers, Allahabad; 1935.
2. Mo S, Hui X, Chu G, Yang X, Wang J, Zheng C, et al. Phaseoloideside E, a novel natural triterpenoid saponin identified from Entada phaseoloides, induces apoptosis in Ec-109 esophageal cancer cells through reactive oxygen species generation. J Pharmacol Sci 2013;122:163-75.
3. Fu XM, Zhang MQ, Shao CL, Li GQ, Bai H, Dai GL, et al. Chinese marine materia medica resources: status and potential. Mar Drugs 2016;14:46.
4. Nanjing University of Traditional Chinese Medicine. Dictionary of Chinese Materia Medica. Shanghai: Shanghai Science and Technology Publishing House; 2006. p. 3563.
5. Entada phaseoloides, Australian Tropical Rainforest Plant; 2020.
6. Siddhuraju P, Becker K, Makkar HP. Chemical composition, protein fractionation, essential amino acid potential and antimetabolic constituents of an unconventional legume, Gila bean (Entada phaseoloides Merrill) seed kernel. J Sci Food Agric 2001;82:191-202.
7. Dawane JS, Pandit VA, Rajopadhye BD. Experimental evaluation of the anti-inflammatory effect of topical application of Entada phaseoloides merill seeds as paste and ointment. N Am J Med Sci 2011;3:513-7.
8. Zheng T, Shu GW, Yang ZZ, Mo SS, Zhao Y, Mei Z, et al. Antidiabetic effect of total saponins from Entada phaseoloides (L.) Merr. In type 2 diabetic rats. J Ethnopharmacol 2012;139:814-21.
9. Liu WC. Anti-tumorous saponin extract from Entada phaseoloides. Chem Abst 1972;76:131-47.
10. Shankar R, Tripathi AK, Anku G, Neyaz S, Rawat MS. Indigenous medicinal plants of northeast india in human health: literary note. J Drug Res Ayur Sci 2017;2:104-17.
11. Barua CC, Buragohain L, Purkayastha A, Saikia B, Babu KS, Kumari GS, et al. Entada phaseoloides attenuates scopolamine induced memory impairment, neuroinflammation and neuro-degeneration via BDNF/TRKb/NF?B p65 pathway in radial arm maze. Int J Pharm Pharm Sci 2018;10:29-38.
12. Barua CC, Buragohain L, Purkayastha A, Rizavi H, Gogoi SB, Rahman F, et al. Effect of seeds of Entada phaseoloides on chronic restrain stress in mice. J Ayur Integrat Med 2018;1-7. Doi:10.1016/j.jaim.2018.02.140.
13. Singh O, Ali M, Akhtar NJ. Phenolic acid glucosides from the seeds of Entada phaseoloides merill. Asian Nat Prod Res 2011;13:682-7.
14. Ikegami F, Sekine T, Duangteraprecha S, Matsushita N, Matsuda N, Ruangrungsi N, et al. Entadamide C-a sulphur-containing amide from Entada phaseoloides. Phytochemistry 1989;28:881-2.
15. Ikegami F, Ohmiya S, Ruangrungsi N, Sakai S, Murakoshi I. Entadamide B, a second new sulfur-containing amide from Entada phaseoloides. Phytochemistry 1987;26:1525-6.
16. Ionkova I, Momekov G, Proksch P. Effects of cycloartane saponins from hairy roots of Astragalus membranaceus Bge., on human tumor cell targets. Fitoterapia 2010;81:447-51.
17. Okada Y, Shibata S, Ikekawa T, Javellana AMJ, Kamo O. Entada saponin-III, a saponin isolated from the bark of Entada phaseoloides. Phytochemistry 1987;26:2789-96.
18. Ahmad F, Khan RA, Rasheed S. Preliminary screening of methanolic extracts of Celastrus paniculatus and Tecomella undulata for analgesic and anti-inflammatory activities. J Ethnopharmacol 1994;42:193-8.
19. Stasi LCD, Costa M, Mendacolli SLJ, Kirizawa M, Gomes C, Trolin G, et al. Screening in mice of some medicinal plants used for analgesic purposes in the state of Sao Paulo. J Ethnopharmacol 1988;24:205-11.
20. Marmitt DJ, Bitencourt S, Silva ADCE, Goettert MI, Rempel C. Medicinal plants of renisus with analgesic activity. J Crit Rev 2016;3:1-4.
21. Dawane JS, Pandit VA, Rajopadhye BD. Anti-nociceptive effect of Entada phaseoloides seeds formulation after topical application in arthritic wistar rats. J Clin Diag Res 2013;7:2744-6.
22. Dawane JS, Pandit VA, Rajopadhye BD. Experimental evaluation of anti-inflammatory effect of topical application of Entada phaseoloides seeds as paste and ointment. N Am J Med Sci 2011;3:513-7.
23. Gupta R, Rathi B, Thakurdesai P, Bodhankar S. Anti-inflammatory and analgesic effects of methanolic extract of Entada phaseoloides seeds. Cell Tissue Res 2006;6:609-13.
24. Lahoti A, Kalra BS, Tekur U. Evaluation of the analgesic and anti-inflammatory activity of fixed-dose combination: non-steroidal anti-inflammatory drugs in experimental animals. Indian J Dent Res 2014;25:551-4.
25. Turner RA. Screening methods in pharmacology: analgesic agent. Vol. 13. Academic Press: London, UK; 1965.
26. Koster R, Anderson M, De J. Acetic acid for analgesic screening. Fed Proc 1959;18:412-7.
27. Eddy NB, Leimbach DJ. Synthetic analgesic dithienyl butanyl and dithinyl butylamines. J Pharmacol Exp Ther 1953;107:385-93.
28. Turner R, Hebborn P. Screening methods in pharmacology: analgesic agent. 2nd ed. London: Academic Press; 1965.
29. Verma A, Jana GK, Chakraborty R, Sen S, Sachan S, Mishra A. Analgesic activity of various leaf extracts of Saraca indica linn. Pharm Lett 2010;2:352-7.
30. Dong Y, Shi H, Yang H, Peng Y, Wang M, Li X. Antioxidant phenolic compounds from the stems of Entada phaseoloides. Chem Biodivers 2012;9:68-79.
31. Naima J, Islam MR, Proma NM, Afrin SR, Hossain MR, Hossain MK. Phytochemical screening and antinociceptive activity of Mimosa diplotricha leaves. Int J Pharma Sci Res 2019;10:3691-6.
32. Hameed JB, Ramadhan UH. Xanthine oxidase inhibitory, antihyperuricemic, anti-inflammatory, antinociceptive activity of ?-lipoic acid in gouty arthritis model. Asian J Pharm Clin Res 2018;11:483-7.
33. Gawade PS. Acetic acid induced painful endogenous infliction in writhing test on mice. J Pharmacol Pharmacother 2012;3:348.
34. Kumar MA, Jitendra J. Anti-inflammatory and analgesic activity of bark extract of Pterospermum acerifolium. Int J Curr Pharm Res 2009;1:32-7.
35. Sunday OO, Om U. Peripheral and central antinociceptive activities of the crude methanolic extract and fractions of Momordica balsamina linn. Asian J Pharm Clin Res 2010;3:15-9.
36. Carlsson KH, Jurna I. Depression by flupirtine a novel analgesic agent of motor and sensory responses of the nociceptive system in the rat spinal cord. Eur J Pharmacol 1987;143:89-9.
37. Chen YF, Li N, Jiao YL. Antinociceptive activity of petroleum ether fraction from the MeOH extracts of Paederia scandens in mice. Phytomedicine 2008;15:427-36.
38. Calixto JB, Beirith A, Ferreira J, Santos ARS. Naturally occurring anti-nociceptive substances from plants. Phytother Res 2000;14:401-18.
39. Berkowitz BA. The relationship of pharmacokinetics to pharmacological activity: morphine, methadone and naloxone. Clin Pharmacokinet 1976;1:219-30.
40. Ainsah O, Nabishah, Osman CB, Khalid BAK. Effects of naloxone, glycerrhyzic acid, dexamethasone and deoxycorticosterone in repetitive stress. Clin Exp Pharmacol Physiol 1999;26:433-7.
41. Kamo KK, Mahlberg PG. Morphin an alkaloids: biosynthesis in plant (Papaver spp.) tissue cultures in medicinal and aromatic plants I of biotechnology in agriculture and forestry. Springer 1988;4:251-63.
42. Kaithwas G, Mukherjee A, Chaurasia AK, Majumdar DK. Anti-inflammatory, analgesic and antipyretic activities of Linum usitatissimum L. (flax seed/linseed) fixed oil. Indian J Exp Biol 2011;49:932-8.
43. Moncada S, Gryglewski R, Bunting S. An enzyme isolated from arteries transforms prostaglandin endoperoxides to an unstable substance that inhibits platelet aggregation. Nature 1976;263:663-5.
44. Whittle BJ, Smith NK, Moncada S. Actions of prostacyclin (PGI2) and its product, 6-oxo-PGF1alpha on the rat gastric mucosa in vivo and in vitro. Prostaglandins 1978;15:955-67.
45. Lee KM, Kang BS, Lee HL, Son SJ, Hwang SH, Kim DS, et al. Spinal NF?B activation induces COX-2 upregulation and contributes to inflammatory pain hypersensitivity. Eur J Neurosci 2004;9:3375-81.
46. Opal SM, DePalo HA. Anti-inflammatory cytokines. Chest 2000;117:1162-72.
47. Clarke CJP, Hales A, Hunt A. IL-10 mediated suppression of TNF-? production is independent of its ability to inhibit NF?B activity. Eur J Immunol 1998;28:1719-26.
48. Opal SM, Wherry JC, Grint P. Interleukin-10:potential benefits and possible risks in clinical infectious diseases. Clin Infect Dis 1998;27:1497-507.
49. Lotta T, Vidgren J, Tilgmann C, Ulmanen I, Melén K, Julkunen I, et al. Kinetics of human soluble and membrane bound catechol O-methyltransferase: a revised mechanism and description of the thermolabile variant of the enzyme. Biochem 1995;34:4202-10.
50. Zubieta JK, Heitzeg MM, Smith YR, Bueller JA, Xu K, Xu Y, et al. COMT val158met genotype affects mu opioid neurotransmitter responses to a pain stressor. Science 2003;299:1240-3.
51. Khasar SG, McCarter G, Levine JD. Epinephrine produces a ?-adrenergic receptor-mediated mechanical hyperalgesia and in vitro sensitization of rat nociceptors. J Neurophysiol 1999;81:1104-12.
52. Sadhasivam S, Chidambaran V. Pharmacogenomics of opioids and perioperative pain management. Pharmacogenomics 2012;13:1719-40.
53. Seibert K, Zhang Y, Leahy K, Hauser S, Masferrer J, Perkins W, et al. Pharmacological and biochemical demonstration of the role of cyclooxygenase 2 in inflammation and pain. Proc Nat Acad Sci USA 1994;91:12013-7.
54. Yamagata K, Andreasson KI, Kaufmann WE, Barnes CA, Worley PF. Expression of a mitogen-inducible cyclooxygenase in brain neurons: regulation by synaptic activity and glucocorticoids. Neuron 1993;11:371-86.
55. Komhoff M, Grone HJ, Klein T. Localization of cyclooxygenase-1 and-2 in adult and fetal human kidney: implication for renal function. Am J Physiol 1997;272:460-8.
56. Vane JR, Bakhle YS, Botting RM. Cyclooxygenases 1 and 2. Ann Rev Pharmacol Toxicol 1998;38:97-120.
57. McCleskey EW, Gold MS. Ion channels of nociception. Ann Rev Physiol 1999;61:835-56.
58. Woolf CJ, Salter MW. Neuronal plasticity-increasing the gain in pain. Science 2000;288:1765-9.
59. Sirois JD, Simmons L, Richards J. Hormonal regulation of messenger ribonucleic acid encoding a novel isoform of prostaglandin endoperoxide H synthase in rat pre ovulatory follicles: induction in vivo and in vitro. J Biol Chem 1992;267:11586-92.
60. Ehrich EW, Dallob A, De LepeleireI. Characterization of rofecoxib as cyclooxygenase-2 isoform inhibitor and demonstration of analgesia in the dental pain model. Clin Pharmacol Ther 1999;65:336-47.
61. Hutchinson MR, Coats BD, Lewis SS. Pro-inflammatory cytokines oppose opioid-induced acute and chronic analgesia. Brain Behav Immun 2008;22:1178-89.
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How to Cite
BARUA, C. C., S. M. A. A. BARBHUIYA, L. BURAGOHAIN, A. KAKATI, and A. G. BARUA. “INVESTIGATING THE ROLE OF NF-κB, COX-1, COX-2, COMT, IL-10, IL-6 AND TNF-α IN MODULATING ANTI-NOCICEPTIVE ACTIVITY OF METHANOLIC EXTRACT OF ENTADA PHASEOLOIDES”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 12, no. 8, July 2020, pp. 172-9, doi:10.22159/ijpps.2020v12i8.38104.
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